质粒名称: | pcDNA3.1(+) CircRNA Mini Vector |
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出品公司: | 爱迪基因 |
目录编号: | 60648 |
存储实验室: | Jeremy Wilusz |
载体骨架基本信息 |
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载体名称: | pcDNA3.1(+) |
载体出品公司: | Life Technologies |
载体抗性: | 氨苄霉素 |
空载体大小: | 5428 bp |
完整质粒大小: | 5607 bp |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | ZKSCAN1 |
物种来源: | H. sapiens (human) |
插入大小: | 199 bp |
启动子 | CMV |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | EcoRI(未破坏) |
3' 克隆位点: | NotI(未破坏) |
5' 测序引物 | PCMV For |
3' 测序引物 | bGH PolyA Rev |
文献引用方法 |
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材料和方法部分: | pcDNA3.1(+) CircRNA Mini Vector was a gift from Jeremy Wilusz (爱迪基因 plasmid # 60648) |
参考文献部分: | Short intronic repeat sequences facilitate circular RNA production. Liang D, Wilusz JE. Genes Dev. 2014 Oct 3. pii: gad.251926.114. 10.1101/gad.251926.114 PubMed 25281217 |
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LOCUS pcDNA3.1(+) CircRNA Mini Vector 5607 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION Expression plasmid for expressing circular RNAs of a desired sequence. ACCESSION . VERSION . KEYWORDS pcDNA3.1(+) CircRNA Mini Vector SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5607) AUTHORS Liang D, Wilusz JE TITLE Short intronic repeat sequences facilitate circular RNA production. JOURNAL Genes Dev. 2014 Oct 3. pii: gad.251926.114. PUBMED 25281217 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|pcDNA3.1(+) CircRNA Mini Vector| FEATURES Location/Qualifiers source 1..5607 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1201..1218) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1207..1431 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1477..1905 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1564..1583) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1774..1795 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1914..1934) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1919..2248 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2099..2234 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2161..2180 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2315..3109 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2369..2388) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2979..2998 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3283..3404 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3320..3339) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3374..3393 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3453..3469) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3453..3469) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3466..3488) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3477..3493 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3501..3531) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3546..3567 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3684..3701) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3855..4440) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3935..3954) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4611..5471) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5234..5253 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5472..5576) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcaaa 961 gtgctgagat tacaggcgtg agccaccacc cccggcccac tttttgtaaa ggtacgtact 1021 aatgactttt tttttatact tcaggatatc atcgataccg gtttaattaa cacgtgggta 1081 accgttaacc cgcggaggta agaagcaagg aaaagaatta ggctcggcac ggtagctcac 1141 acctgtaatc ccagcagcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag 1201 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 1261 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 1321 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 1381 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 1441 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 1501 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 1561 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 1621 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 1681 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 1741 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 1801 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 1861 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 1921 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 1981 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 2041 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 2101 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 2161 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 2221 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg 2281 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc 2341 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat 2401 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt 2461 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg 2521 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag 2581 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc 2641 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc 2701 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga 2761 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga 2821 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg 2881 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg 2941 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc 3001 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc 3061 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg 3121 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc 3181 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc 3241 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct 3301 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 3361 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg 3421 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt 3481 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 3541 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 3601 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 3661 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 3721 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 3781 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 3841 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 3901 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 3961 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 4021 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 4081 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 4141 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 4201 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 4261 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg 4321 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 4381 gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa 4441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa 4501 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa 4561 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc 4621 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga 4681 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca 4741 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc 4801 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat 4861 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc 4921 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt 4981 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc 5041 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg 5101 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt 5161 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg 5221 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga 5281 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg 5341 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg 5401 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt 5461 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc 5521 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 5581 tttccccgaa aagtgccacc tgacgtc //
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